TOOLShinning Dual Luciferase Reporter Assay Kit
TOOLShinning Dual Luciferase Reporter Assay Kit detects gene regulation by transfecting cells with a reporter plasmid and measuring the fluorescence intensity of Luciferin substrate to reflect the level of Luciferase expression. To achieve dual luciferase reporter gene detection, firefly luciferase is detected with Luciferin as a substrate, and renilla luciferase is detected with coelenterazine as a substrate, while inhibiting the activity of Firefly luciferase. Dual Luciferase Reporter Assay Kit can detect the expression of Luciferase regulated by gene elements sensitively and efficiently. Usually, the transcriptional regulatory element is cloned upstream of Firefly luciferase, or the 3'-UTR regulatory region is cloned downstream of firefly luciferase. The transfected cells are induced by corresponding stimulator and lysed to determine the luciferase activity. The stimulatory-inducing effect of the regulatory elements is evaluated by luciferase activity. Renilla luciferase acts as an internal reference for correcting transfection efficiency to eliminate differences in cell number and transfection efficiency between wells. Firefly luciferase catalyzes the emission of Luciferin at 560 nm, and renilla luciferase catalyzes the emission of coelenterazine at 465 nm.
Sensitive: measure Green Renilla and red firefly luciferase activities in the same sample
Efficient: addition of the Stop/go Buffer can inhibit 99% of the Firefly luciferase activity
Concurrent: Detect filter-based, wavelength-separated two luciferase activities